Various media for growing eukaryotic cells in culture are described in the literature and are commercially available. Cell culture media functions to provide the cells with a suitable pH and osmolality and nutrients essential for cell survival, growth and protein expression.
Examples of some of some common basal culture media are RPMI Media 1640, Medium 199, Minimal Essential Medium (MEM) medium (a “minimal” medium for growth of attached mammalian cells), Leibovitz medium for growth in absence of CO2, Dulbecco's Modified Eagle's Medium and Ham's F12 Medium. The various media are distinguished from one another in that they contain different components in precise amino acids, vitamins, organic salts, trace elements, and other organic compounds which promote the growth of (and protein expression by) the cultured cells.
The development of an optimal cell culture medium is of significant importance, because changes to various components can lead to unexpected improvements in cell growth, increased growth rates, growth to high cell densities, improvements in controlling the stage and amount of cell differentiation, increased protein secretion, increased phenotypic and genetic stability, and elimination of senescence for many cell types, all of which are consequential properties when producing recombinant proteins on a commercial scale.
Preparation of cell culture media, in particular in a commercial setting, is complex, and usually requires the stocking, transfer, preparation and storage of multiple stock solutions and powders and batching in a sequential manner. The large number of process steps and components in media preparation reduce efficiency and increase costs; they can also introduce variability, which in turn can impact the growth of cells and protein production. Hydration is also a significant obstacle to overcome, because as more and more components are added to a media formulation, the more difficult and less predictable it becomes to completely hydrate and dissolve the components to achieve a homogeneous mixture and an optimal osmolality, due to the richness and complexity of the composition, and interactions amongst the various components. For this reason, some components are often hydrated separately in complex media formulations. However, this results in increased cost and complexity during production.
Accordingly, there remains a need in the art for improved/further compounded dry media powder (DMP) formulations and methods of making the same in order to reduce complexity and lower costs while maintaining the correct composition in the fully hydrated liquid media as well as similar cell culture performance and recombinant protein production.